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A procedure for purifying jack bean urease for clinical use.
1Department of Agricultural Chemistry, National Taiwan University, Taipei, R.O.C.
Proceedings of the National Science Council, Republic of China. Part B, Life Sciences
|October 1, 1989
Summary
A simplified jack bean urease purification method yields a highly pure enzyme suitable for analytical and clinical applications, achieving a 14.7-fold increase in purity.
Area of Science:
- Biochemistry
- Enzyme Purification
Background:
- Urease is a crucial enzyme for various analytical and clinical assays.
- Efficient purification of urease from natural sources like jack bean is essential for its practical application.
Purpose of the Study:
- To develop a simple and effective purification procedure for jack bean urease.
- To obtain urease with purity suitable for analytical use.
Main Methods:
- Extraction using 20% acetone with EDTA and 2-mercaptoethanol.
- Heat treatment, acid precipitation (pH 5.4), neutralization, and lyophilization.
- Analysis of purity and activity using gel filtration and polyacrylamide gel electrophoresis.
Main Results:
- Achieved a 14.7-fold increase in urease purity.
- Obtained 63% recovery of activity and a yield of 6.75 g/kg bean seeds.
- Specific activity was 411 units/mg protein; molecular weight estimated at 480,000, with two activity bands observed.
Conclusions:
- The developed purification method is simple and practical for preparing analytical-grade urease from jack bean.
- The purified urease is suitable for clinical analysis, such as blood urea nitrogen determination.